This application claims priority under 35 U.S.C. xc2xa7119 to Korean patent applications KR 1998-0060266 filed Dec. 29, 1998 and KR 1999-0033490 filed Aug. 14, 1999.
The present invention relates to thrombin inhibitors that are useful as anticoagulants. In particular, this invention relates to peptide derivatives having high antithrombotic activity and high oral bioavailability.
Thrombosis is characterized by excessive blood clotting. The condition plays a significant role in cardiovascular and related diseases, and thrombotic events underlie a significant proportion of the mortality and morbidity associated with cardiovascular disease. Thrombosis can cause a range of disease states which are characterized by the location of the blood vessel in which the clot is formed.
Thrombin is a trypsin-like serine protease that plays a key role in the blood coagulation cascade by catalyzing the conversion of fibrinogen to insoluble fibrin. This enzyme also activates Factor V and Factor VIII for its own production and potently activates platelets as well. Therefore, thrombin has long been recognized as a central regulator in thrombosis and hemostasis, and its inhibition has become a major therapeutic target in the treatment of cardiovascular diseases such as myocardial infarction, unstable angina, deep vein thrombosis and pulmonary embolism. Indirect thrombin inhibitors such as heparin and warfarin (coumarin) have been used as antithrombotic therapies with, however, several limitations. Heparin demonstrates low bioavailability and is associated with side effects such as bleeding problems, moreover, it is not able to inhibit clot-bound thrombin. Warfarin is an effective oral anticoagulant but it has a narrow therapeutic window and also requires patient monitoring. A natural protein inhibitor, hirudin, has been associated with bleeding complications.
Most of the low molecular weight thrombin inhibitors are broadly based upon peptides or peptidomimetic templates which operate by a direct mechanism of action against the target enzyme. Early examples are tripeptidic aldehydes such as D-Phe-Pro-Arg-H and Me-D-Phe-Pro-Arg-H that have been reported to be effective thrombin inhibitors (Bajusz et al. J. Med. Chem. 1990, 33, 1729).
Recently, D-Phe-Pro-Agmatine and its derivatives have been described as thrombin inhibitors in U.S. Pat. No. 4,346,078 and WO93/11152 (agmatine=1-amino-4-guanidinobutane). These compounds are different from the earlier tripeptidic compounds in that the agimatine compounds lack a carbonyl moiety found in similar compounds containing an Arg side chain.
More recently, certain tripeptidic thrombin inhibitors in which 4-amidinobenzylamaine was incorporated at the P1 position in place of agmatine have been disclosed (WO 94/29336). These amidine-based compounds have been reported to possess good antithrombotic activity (WO 95/23609). However, this class of compounds has generally poor or low oral bioavailability.
Certain thrombin inhibitors bearing the unique amino acid D-diphenylalanine at P3 position have been disclosed (WO 93/11152, U.S. Pat. No. 5,510,369, WO 97/15190). These compounds have been reported to have higher potency against thrombin compared to the corresponding D-phenylalanine alalogs (J. Med. Chem. 1992, 35, 3365; J. Med. Chem. 1997, 40, 830). In addition, some of this class of compounds exhibited good oral bioavailability (J. Med. Chem. 1997, 40, 3687; J. Med. Chem. 1997, 40, 3726).
In certain thrombin inhibitors and Factor Xa inhibitors, the amidinothiophene groups have been shown to be better para-benzamidine surrogates (WO 95/23609, WO98/24784, Bioorg. Med. Chem. Lett. 1998, 8,1683). In addition, 2,5-thiophene and other 5-membered heterocyclic moieties have effectively served as a para-phenylene isostere in the inhibitors of other drug-targeting enzymes such as thymidylate synthase and glycinamide riobonuceotide formyltransferase (J. Med. Chem. 1991, 34, 1594; Cancer Research 1994, 54,1021; WO 97/41115).
Therefore, there is a need in the art for thrombin inhibitors which have improved oral bioavailability and stablility as compared to those described supra. We have found that the compounds of the present invention, as defined below, are potent inhibitors of thrombin in vitro and in vivo. In particular, certain compounds of this invention exhibit high bioavailability after oral administration.
The present invention relates to compounds falling within formula I below which modulate and/or inhibit the serine protease thrombin, as well as to acceptable prodrugs, pharmaceutically active metabolites, and pharmaceutically acceptable salts thereof (such compounds, prodrugs, metabolites and salts are collectively referred to as xe2x80x9cagentsxe2x80x9d). The invention is also directed to pharmaceutical compositions containing such agents and their therapeutic use in treating diseases mediated by thrombin, such as myocardial infarction, unstable angina, deep vein thrombosis and pulmonary embolism, as well as other disease states associated with blood clotting and associated clotting factors.
In one general aspect, the invention relates to thrombin inhibitors of the Formula I: 
and pharmaceutically acceptable salts thereof
wherein:
n is 1, 2, or 3,
A is hydrogen, alky, C3-7 cycloalkyl, aryl, xe2x80x94SO2R1, xe2x80x94SO3R1, xe2x80x94COR1, xe2x80x94CO2R2, xe2x80x94PO(R1)2, xe2x80x94PO(OR1)2, xe2x80x94(CH2)mCO2R1, (CH2)mSO2R1, xe2x80x94(CH2)mSO3R1, or xe2x80x94(CH2)mPO(OR1)2,
wherein:
R1 is hydrogen, C1-6 alkyl, C3-7 cycloalkyl, aryl, xe2x80x94(CH2)maryl, or xe2x80x94NR3R4, and
R2 is C1-6 alkyl, C3-7 cycloalkyl, aryl, xe2x80x94(CH2)maryl, or alkenyl,
m is 1, 2, or 3,
wherein:
aryl is unsubsituted or substituted phenyl or 5-6 membered aromatic heterocyclic ring,
R3 and R4 are independently hydrogen, C1-6 alkyl, or C3-7 cycloalkyl;
B is hydrogen or C1-6 alkyl;
C and D are independently hydrogen, unsubsituted or substituted phenyl with one or two substituents selected from C1-4 alkyl, C1-4 alkoxy, CF3, methylenedioxy, halogen, hydroxy, or xe2x80x94NR3R4, C3-7 cycloalkyl, or a 5-6 membered heterocyclic ring system which may be saturated or unsaturated, and which consists of carbon atoms and 1-3 heteroatoms selected from the group consisting of N, O, and S; 
xe2x80x83wherein
X is S, O, or NR5,
Y and Z are independently N or CR6,
wherein
R5 is hydrogen or C1-4 alkyl and
R6 is hydrogen, halogen, CF3 or C1-4 alkyl; and
F is xe2x80x94C(NH)N(R7)2, xe2x80x94C(NH2)NN(R7)2, xe2x80x94C(NH2)NOH, or xe2x80x94CH2NH(R7)2 
wherein R7 is the same or different,
R7 is hydrogen, C1-4 perfluoroalkyl or C1-4 alkyl.
The invention also relates to pharmaceutical compositions each comprising: an effective amount of an agent selected from compounds of Formula I and pharmaceutically acceptable salts, pharmaceutically active metabolites, and pharmaceutically acceptable prodrugs thereof; and a pharmaceutically acceptable carrier or vehicle for such agent. The invention further provides methods of treating cardiovascular diseases such as myocardial infarction, unstable angina, deep vein thrombosis and pulmonary embolism, as well as other disease states associated with excess thrombin.
The inventive compounds of Formula I are useful for mediating the activity of trypsin-like serine proteases. More particularly, the compounds are useful as anti-coagulant agents and as agents for modulating and/or inhibiting the activity of trypsin-like serine proteases, thus providing treatments for thrombosis and other cardiovascular diseases such as myocardial infarction, unstable angina, deep vein thrombosis and pulmonary embolism.
The terms and abbreviations used in the instant disclosure have their normal meanings unless otherwise designated.
As used in the present application, the following definitions apply:
In accordance with a convention used in the art, 
is used in structural formulas herein to depict the bond that is the point of attachment of the moiety or substituent to the core or backbone structure.
Where chiral carbons are included in chemical structures, unless a particular orientation is depicted, both stereoisomeric forms are intended to be encompassed.
An xe2x80x9calkyl groupxe2x80x9d is intended to mean a straight or branched chain monovalent radical of saturated and/or unsaturated carbon atoms and hydrogen atoms, such as methyl (Me), ethyl (Et), propyl, isopropyl, butyl (Bu), isobutyl, t-butyl (t-Bu), ethenyl, pentenyl, butenyl, propenyl, ethynyl, butynyl, propynyl, pentynyl, hexynyl, and the like, which may be unsubstituted (i.e., containing only carbon and hydrogen) or substituted by one or more suitable sustituents as defined below (e.g., one or more halogens, such as F, Cl, Br, or I, with F and Cl being preferred).
A xe2x80x9clower alkyl groupxe2x80x9d is intended to mean an alkyl group having from 1 to 8 carbon atoms in its chain.
A xe2x80x9ccycloalkyl groupxe2x80x9d is intended to mean a non-aromatic monovalent monocyclic, bicyclic, or tricyclic radical containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 carbon ring atoms, each of which may be saturated or unsaturated, and which may be unsubstituted or substituted by one or more suitable substituents as defined below, and to which may be fused one or more heterocycloalkyl groups, aryl groups, or heteroaryl groups, which themselves may be unsubstituted or substituted by one or more substituents.
A xe2x80x9cheterocycloalky groupxe2x80x9d is intended to mean a non-aromatic monovalent monocyclic, bicyclic, or tricyclic radical, which is saturated or unsaturated, containing 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, which includes 1, 2, 3, 4, or 5 heteroatoms selected nitrogen, oxygen, and sulfur, where the radical is unsubstituted or substituted by one or more suitable substituents as defined below, and to which may be fused one or more cycloalkyl groups, aryl groups, or heteroaryl groups, which themselves may be unsubstituted or substituted by one or more suitable substituents.
An xe2x80x9caryl groupxe2x80x9d is intended to mean an aromatic monovalent monocyclic, bicyclic, or tricyclic radical containing 6, 10, 14, or 18 carbon ring atoms, which may be unsubstituted or substituted by one or more suitable substituents as defined below, and to which may be fused one or more cycloalkyl groups, heterocycloalkyl groups, or heteroaryl groups, which themselves may be unsubstituted or substituted by one or more suitable substituents. Thus, the term xe2x80x9caryl groupxe2x80x9d includes a benzyl group (Bzl)., or tricyclic radical containing 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 ring atoms, including 1, 2, 3, 4, or 5 heteroatoms selected from nitrogen, oxygen, and sulfur, which may be unsubstituted or substituted by one or more suitable substituents as defined below, and to which may be fused one or more cycloalkyl groups, heterocycloalkyl groups, or aryl groups, which themselves may be unsubstituted or substituted by one or more suitable substituents.
A xe2x80x9cheterocyclexe2x80x9d is intended to mean a heteroaryl or heterocycloalkyl group (each of which, as defined above, are optionally substituted).
An xe2x80x9cacyl groupxe2x80x9d is intended to mean a xe2x80x94C(O)xe2x80x94R radical, where R is a substituent as defined below.
A xe2x80x9cthioacyl groupxe2x80x9d is intended to mean a xe2x80x94C(S)xe2x80x94R radical, where R is a substituent as defined below.
A xe2x80x9csulfonyl groupxe2x80x9d is intended to mean a xe2x80x94SO2R radical, where R is a substituent as defined below.
A xe2x80x9chydroxy groupxe2x80x9d is intended to mean the radical xe2x80x94OH.
An xe2x80x9camino groupxe2x80x9d is intended to mean the radical xe2x80x94NH2.
An xe2x80x9calkylamino groupxe2x80x9d is intended to mean the radical xe2x80x94NHRa, where Ra is an alkyl group.
Axe2x80x9cdialkylamino groupxe2x80x9d is intended to mean the radical xe2x80x94NRaRb, where Ra and Rb are each independently an alkyl group.
An xe2x80x9calkoxy groupxe2x80x9d is intended to mean the radical xe2x80x94ORa, where Ra is an alkyl group. Exemplary alkoxy groups include methoxy, ethoxy, propoxy, and the like.
An xe2x80x9calkoxycarbonyl groupxe2x80x9d is intended to mean the radical xe2x80x94C(O)ORa, where Ra is an alkyl group.
An xe2x80x9calkylsulfonyl groupxe2x80x9d is intended to mean the radical xe2x80x94SO2Ra, where Ra is an alkyl group.
An xe2x80x9calkylaminocarbonyl groupxe2x80x9d is intended to mean the radical xe2x80x94C(O)NHRa, where Ra is an alkyl group.
A xe2x80x9cdialkylaminocarbonyl groupxe2x80x9d is intended to mean the radical xe2x80x94C(O)NRaRb, where Ra and Rb are each independently an alkyl group.
A xe2x80x9cmercapto groupxe2x80x9d is intended to mean the radical xe2x80x94SH.
An xe2x80x9calkylthio groupxe2x80x9d is intended to mean the radical xe2x80x94SRa, where Ra is an alkyl group.
A xe2x80x9ccarboxy groupxe2x80x9d is intended to mean the radical xe2x80x94C(O)OH.
A xe2x80x9ccarbamoyl groupxe2x80x9d is intended to mean the radical xe2x80x94(O)NH2.
An xe2x80x9caryloxy groupxe2x80x9d is intended to mean the radical xe2x80x94ORc, where Rc is an aryl group.
A xe2x80x9cheteroaryloxy groupxe2x80x9d is intended to mean the radical xe2x80x94ORd, where Rd is a heteroaryl group.
An xe2x80x9carylthio groupxe2x80x9d is intended to mean the radical xe2x80x94SRc, where Rc is an aryl group.
A xe2x80x9cheteroarylthio groupxe2x80x9d is intended to mean the radical xe2x80x94SRd, where Rd is a heteroaryl group.
A xe2x80x9cleaving groupxe2x80x9d (Lv) is intended to mean any suitable group that will be displaced by a substitution reaction. One of ordinary skill in the art will know that any conjugate base of a strong acid can act as a leaving group. Illustrative examples of suitable leaving groups include, but are not limited to, xe2x80x94F, xe2x80x94Cl, xe2x80x94Br, alkyl chlorides, alkyl bromides, alkyl iodides, alkyl sulfonates, alkyl benzenesulfonates, alkyl p-toluenesulfonates, alkyl methanesulfonates, triflate, and any groups having a bisulfate, methyl sulfate, or sulfonate ion.
Typical protecting groups, reagents and solvents are well known in the art. One skilled in the art would know possible protecting groups, reagents and solvents; these are intended to be within the scope of this invention.
The term xe2x80x9csuitable organic moietyxe2x80x9d is intended to mean any organic moiety recognizable, such as by routine testing, to those skilled in the art as not adversely affecting the inhibitory activity of the inventive compounds. Illustrative examples of suitable organic moieties include, but are not limited to, hydroxyl groups, alkyl groups, oxo groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, acyl groups, sulfonyl groups, mercapto groups, alkylthio groups, alkoxy groups, carboxy groups, amino groups, alkylamino groups, dialkylamino groups, carbamoyl groups, arylthio groups, heteroarylthio groups, and the like.
The term xe2x80x9csubstituentxe2x80x9d or xe2x80x9csuitable substituentxe2x80x9d is intended to mean any suitable substituent that may be recognized or selected, such as through routine testing, by those skilled in the art. Illustrative examples of suitable substituents include hydroxy groups, halogens, oxo groups, alkyl groups, acyl groups, sulfonyl groups, mercapto groups, alkylthio groups, alkyloxy groups, cycloalkyl groups, heterocycloalkyl groups, aryl groups, heteroaryl groups, carboxy groups, amino groups, alkylamino groups, dialkylamino groups, carbamoyl groups, aryloxy groups, heteroaryloxy groups, arylthio groups, heteroarylthio groups, and the like.
The term xe2x80x9coptionally substitutedxe2x80x9d is intended to expressly indicate that the specified group is unsubstituted or substituted by one or more suitable substituents, unless the optional substituents are expressly specified, in which case the term indicates that the group is unsubstituted or substituted with the specified substituents. As defined above, various groups may be unsubstituted or substituted (i.e., they are optionally substituted) unless indicated otherwise herein (e.g., by indicating that the specified group is unsubstituted).
The terms xe2x80x9ccomprisingxe2x80x9d and xe2x80x9cincludingxe2x80x9d are used in an open, non-limiting sense.
A xe2x80x9cpharmaceutically acceptable prodrugxe2x80x9d is a compound that may be converted under physiological conditions or by solvolysis to the specified compound or to a pharmaceutically acceptable salt of such compound.
A xe2x80x9cpharmaceutically active metabolitexe2x80x9d is intended to mean a pharmacologically active product produced through metabolism in the body of a specified compound or salt thereof. Metabolites of a compound may be identified using routine techniques known in the art and their activities determined using tests such as those described herein.
A xe2x80x9cpharmaceutically acceptable saltxe2x80x9d is intended to mean a salt that retains the biological effectiveness of the free acids and bases of the specified compound and that is not biologically or otherwise undesirable. A compound of the invention may possess a sufficiently acidic, a sufficiently basic, or both functional groups, and accordingly react with any of a number of inorganic or organic bases, and inorganic and organic acids, to form pharmaceutically acceptable salt. Exemplary pharmaceutically acceptable salts include those salts prepared by reaction of the compounds of the present invention with a nimeral or organic acid or an inorganic base, such as salts including sulfates, pyrosulfates, bisulfates, sulfites, bisulfites, phosphates, monohydrogenphosphates, dihydrogenphosphates, metaphosphates, pyrophosphates, chlorides, bromides, iodides, acetates, propionates, decanoates, caprylates, acrylates, formates, isobutyrates, caproates, heptanoates, propiolates, oxalates, malonates, succinates, suberates, sebacates, fumarates, maleates, butyne-1,4-dioates, hexyne-1,6-dioates, benzoates, chlorobenzoates, methylbenzoates, dinitrobenzoates, hydroxybenzoates, methoxybenzoates, phthalates, sulfonates, xylenesulfonates, phenylacetates, phenylpropionates, phenylbutyrates, citrates, lactates, y-hydroxybutyrates, glycollates, tartrates, methane-sulfonates, propanesulfonates, naphthalene-1-sulfonates, naphthalene-2-sulfonates, and mandelates.
Some abbreviations that appear in this application are as follows:
Boc: t-butoxycarbonyl
Pro: proline
EDC: 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride
HOBT: 1-hydroxybenzonitrile hydrate
TFA: trifluoroacetic acid
AcOH: acetic acid
DMF: dimethylformamide
EtOAc: ethyl acetate
HCl: hydrochloride
rt: room temperature
TEA: triethylamine
FAB MS: fast atom bombardment mass spectrum
If the inventive compound is a base, the desired pharmaceutically acceptable salt may be prepared by any suitable method available in the art, for example, treatment of the free base with an inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like, or with an organic acid , such as acetic acid, maleic acid, succinic acid, mandelic acid, fumaric acid, malonic acid, pyrovic acid, oxalic acid, glycolic acid, salicylic acid, a pyranosidyl acid, such as glucuronic acid or galacturonic acid, an alpha-hydroxy acid, such as citric acid or tartaric acid, an amino acid, such as aspartic acid or glutamic acid, an aromatic acid, such as benzoic acid or cinnamic acid, a sulfonic acid, such as p-toluenesulfonic acid or ethanesulfonic acid, or the like.
If the inventive compound is an acid, the desired pharmaceutically acceptable salt may be prepared by any suitable method, for example, treatment of the free acid with an inorganic or organic base, such as an amine (primary, secondary or tertiary), an alkali metal hydroxide or alkaline earth metal hydroxide, or the like. Illustrative examples of suitable salts include organic salts derived from amino acids, such as glycine and arginine, ammonia, primary, secondary, and tertiary amines, and cyclic amines, such as piperidine, morpholine and piperazine, and inorganic salts derived from sodium, calcium, potassium, magnesium, manganese, iron, copper, zinc, aluminum and lithium.
In the case of agents that are solids, it is understood by those skilled in the art that the inventive compounds and salts may exist in different crystal or polymorphic forms, all of which are intended to be within the scope of the present invention and specified formulas.
Preferred compounds of the invention include, but are not limited to the following:
1. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
2. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-aminomethyl-2-thienyl)methyl]amide,
3. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
4. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidrazono-2-thienyl)methyl]amide,
5. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-aminomethyl-2-thienyl)methyl]amide,
6. N-benzylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
7. N-t-butoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
8. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
9. N-aminosulfonyl-D-3,4-dichlorophenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
10. N-methoxycarbonyl-D-dicyclohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
11. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thienyl)methyl]amide,
12. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thienyl)methyl]amide,
13. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-aminomethyl-2-thienyl)methyl]amide,
14. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thienyl)methyl]amide,
15. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-thienyl)methyl]amide,
16. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-thienyl)methyl]amide,
17. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[)5-amidrazono-3-thienyl)methyl]amide,
18. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-aminomethyl-3-thienyl)methyl]amide,
19. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-thienyl)methyl]amide,
20. N-cyclohexylsulfamoyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
21. N-allyloxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
22. N-benzylsulfonyl-D-cyclohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
23. N-cyclohexylsulfamoyl-D-cyclohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
24. N-methylsulfamoyl-D-cylohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
25. N-methylsulfonyl-D-cyclohexylglycinyl-L-prolyl-[(5-amidino-3-thienyl)methyl]amide,
26. N-(t-butoxycarbonyl)methyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
27. N-(t-butoxycarbonyl)methyl-D-diphenylalanyl-L-prolyl-[(5-hydroxyamidino-2-thienyl)methyl]amide,
28. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
29. N-methyl-N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
30. N-hydroxysulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
31. N-methylsulfonyl-D-diphenylalanyl-L-azetidine-2-carboxyl-[(5-amidino-2-thienyl)methyl]amide,
32. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-5-methyl-2-thienyl)methyl]amide,
33. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-furanyl)methyl]amide,
34. N-methoxycabonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-furanyl)methyl]amide,
35. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-furanyl)methyl]amide,
36. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thiazolyl)methyl]amide,
37. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-1-methyl-2-pyrrolyl)methyl]amide,
38. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-thienyl)methyl]amide,
39. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thiazolyl)methyl]amide,
40. N-[2-(methoxycarbonyl)ethyl]-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide
41. N-(2-carboxyethyl)-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
42. N-Boc-D-Diphenylalanyl-L-prolyl-[(5-amidrazono-2-thienyl)methyl]amide,
43. D-diphenylalanyl-L-prolyl-[(5-amidrazono-2-thienyl)methyl]amide,
44. N-methoxycabonyl-D-diphenylalanyl-L-azetidine-2-carboxyl-[(5-amidino-2-thienyl)methyl]amide,
45. N-(2-carboxyethyl)-D-diphenylalanyl-L-azetidine-2-carboxyl-[(5-amidino-2-thienyl)methyl]amide,
46. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-hydroxyamidino-2-thienyl)methyl]amide,
47. N-(methoxycarbonyl)methyl-D-diphenylalanyl-L-prolyl-[(5-hydroxyamidino-2-thienyl)methyl]amide,
48. D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
49. N-(3-carboxypropyl)-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
50. N-(MeO)2P(O)-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
51. N-(Me)2P(O)-D-diphenylalanyl-prolyl-[(5-amidino-2-thienyl)methyl]amide,
52. N-(HO)2P(O)-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
53. N-methyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
54. N-phenyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
55. N-[(N, N-diethylcarboxamido) methyl]-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
56. N-[(N,N-diethylcarboxamido)ethyl]-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
57. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thiazolyl)methyl]amide,
58. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-thiazolyl)methyl]amide,
59. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thiazolyl)methyl]amide,
60. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-thiazolyl)methyl]amide,
61. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thiazolyl)methyl]amide,
62. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-thiazolyl)methyl]amide,
63. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thiazolyl)methyl]amide,
64. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-thiazolyl)methyl]amide,
65. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thiazolyl)methyl]amide,
66. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-thiazolyl)methyl]amide,
67. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-thiazolyl)methyl]amide,
68. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-thiazolyl)methyl]amide,
69. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thiazolyl)methyl]amide,
70. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-thiazolyl)methyl]amide,
71. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-thiazolyl)methyl]amide,
72. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-oxazolyl)methyl]amide,
73. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-oxazolyl)methyl]amide,
74. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-oxazolyl)methyl]amide,
75. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-oxazolyl)methyl]amide,
76. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-oxazolyl)methyl]amide,
77. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-oxazolyl)methyl]amide,
78. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-oxazolyl)methyl]amide,
79. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-oxazolyl)methyl]amide,
80. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-oxazolyl)methyl]amide,
81. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-oxazolyl)methyl]amide,
82. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-oxazolyl)methyl]amide,
83. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-oxazolyl)methyl]amide,
84. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-oxazolyl)methyl]amide,
85. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-oxazolyl)methyl]amide,
86. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-oxazolyl)methyl]amide,
87. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(2-amidino-5-oxazolyl)methyl]amide,
88. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-isoxazolyl)methyl]amide,
89. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-isoxazolyl)methyl]amide,
90. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-isoxazolyl)methyl]amide,
91. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-isoxazolyl)methyl]amide,
92. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-isoxazolyl)methyl]amide,
93. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-isoxazolyl)methyl]amide,
94. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-isoxazolyl)methyl]amide,
95. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(2-amidino-4-isoxazolyl)methyl]amide,
96. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-isoxazolyl)methyl]amide,
97. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrazolyl)methyl]amide,
98. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrazolyl)methyl]amide,
99. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrazolyl)methyl]amide,
100. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-pyrrolyl)methyl]amide,
101. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrrolyl)methyl]amide,
102. N-aminosulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-pyrrolyl)methyl]amide,
103. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-pyrrolyl)methyl]amide,
104. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrrolyl)methyl]amide,
105. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-pyrrolyl)methyl]amide,
106. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-pyrrolyl)methyl]amide,
107. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrrolyl)methyl]amide,
108. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-pyrrolyl)methyl]amide,
109. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-pyrrolyl)methyl]amide,
110. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(4-amidino-2-pyrrolyl)methyl]amide,
111. N-carboxymethyl-D-diphenylalanyl-L-prolyl-[(5-amidino-3-pyrrolyl)methyl]amide,
112. N-aminosulfonyl-D-dicyclohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
113. N-methylsulfonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-pyrrolyl)methyl]amide,
114. N-carboxymethyl-D-dicyclohexylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
115. N-aminosulfonyl-D-bis-(para-methoxyphenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
116. N-methoxycarbonyl-D-diphenylalanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
117. N-methoxycarbonyl-D-bis-(para-methoxyphenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
118. N-carboxymethyl-D-bis-(para-methoxyphenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
119. N-aminosulfonyl-D-bis-(para-aminophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
120. N-metylsulfonyl-D-bis-(para-aminophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
121. N-methoxycarbonyl-D-bis-(para-aminophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
122. N-carboxymethyl-D-bis-(para-aminophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
123. N-aminosulfonyl-D-bis-(para-chlorophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
124. N-methylsulfonyl-D-bis-(para-chlorophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
125. N-methoxycarbonyl-D-bis-(para-chlorophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide,
126. N-carboxymethyl-D-bis-(para-chlorophenyl)alanyl-L-prolyl-[(5-amidino-2-thienyl)methyl]amide.
Compounds of the present invention can be prepared according to the general procedure outlined below:
As exemplified by Example 1 and Example 8 (Scheme 1), a protected amino acid such as N-Boc-D-diphenylalnine is coupled to proline methyl ester using a couplng agent such as EDC and HOBT. The resultant dipeptide is treated with a strong acid such as hydrochloric acid gas or trifluoroacetic acid to remove the t-butoxycarbonyl (Boc) protecting group. The resultant free amine is reacted with a sulfonylating reagent such as sulfamoyl chloride and a base such as triethylamine. Carbamate-containing compounds are prepared using chloroformates. The product is then hydrolyzed with base such as lithium hydroxide, and the resultant acid is coupled to the desired amine such as 5-(aminomethyl)thiophene-2-carbonitrile. The coupled product is converted to the amidine by a three-step sequence involving sequential treatment with hydrogen sulfide, methyl iodide, and ammonium acetate. The amidrazone compound is obtained by treatment with hydrazine in place of ammonium acetate in the final step. The nitrile intermediate is also converted to the methylamine by catalytic hydrogenation in the presence of a strong acid such as hydrochloric acid. 
An alternative route, as depicted in Scheme 2, is to hydrolyze the Boc-protected dipeptide before functionalizing the amino group and then couple the resultant acid with the desired amine. The protecting group of the coupling product is removed and the free amine is then sulfonylated. 
N-Boc-D-diphenylalnine can be coupled directly to the amine-coupled proline as exemplified by Example 32 (Scheme 3). The product is then deprotected and subsequently sulfonylated 
Another method for synthesizing compounds of the invention, particularly N-carboxyalkyl substituted compounds, is as exemplified by Example 28 and Example 29 (Scheme 4). The free amino containing compound reacts with an alkylating agent such as t-butylbromoacetate and a base such as diisopropylethylamine (DIPEA). The resulting compound is treated with hydroxylamine hydrochloride in the presence of base such as sodium carbonate, and the resultant amidoxime is catalytically hydrogenated in the presence of acetic anhydride to produce the amidine. The t-butyl group is removed with acid such as trifluoroacetic acid and hydrochloric acid to give the product. 
Amide coupling used to form the compounds of this invention are typically performed by the carbodiimide method with reagents such as dicyclohexylcarbodiimide (DCC) or 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC). Other method for forming the amide or peptide bond include, but not limited to the synthetic routes via an acid chloride, azide, mixed anhydride or activated ester. The addition and removal of one or more protecting groups are typical practice. Methods for suitable protection and deprotection are provided in xe2x80x9cProtective Groups in Organic Synthesisxe2x80x9d, 3rd Edition, by T. W. Green and Peter G. M. Wuts (1999), John Wiley and Sons, Inc., publishers.
The amide coupling reactions are carried out in an inert organic solvent such as dimethylformamide, dimethylacetamide, tetrahydrofuran, dichloromethane, chloroform, and like common solvents or a mixture of such solvents.
Compounds that potently regulate, modulate, or inhibit the conversion of fibrinogen to fibrin via the enzyme thrombin, and therefore inhibit thrombosis and clotting are desirable and represent preferred embodiments of the present invention. The present invention is further directed to methods of modulating trypsin-like serine protease activity, for example in mammalian tissue, by administering an inventive agent. The activity of the inventive compounds as modulators of trypsin-like serine protease activity, such as the activity of thrombin, may be measured by any of the methods available to those skilled in the art, including in vivo and/or in vitro assays. These properties may be assessed, for example, by using one or more of the biological testing procedures set out in the examples below.
The active agents of the invention may be formulated into pharmaceutical compositions as described below. Pharmaceutical compositions of this invention comprise an effective modulating, regulating, or inhibiting amount of a compound of Formula I and an inert, pharmaceutically acceptable carrier or diluent. In one embodiment of the pharmaceutical compositions, efficacious levels of the inventive agents are provided so as to provide therapeutic benefits involving modulation of tyrpsin-like serine proteases. By xe2x80x9cefficacious levelsxe2x80x9d is meant levels in which the effects of tyrpsin-like serine proteases like thrombin are, at a minimum, regulated. These compositions are prepared in unit-dosage form appropriate for the mode of administration, e.g., parenteral or oral administration.
The compositions of the invention may be manufactured in manners generally known for preparing pharmaceutical compositions, e.g., using conventional techniques such as mixing, dissolving, granulating, dragee-making, levigating, emulsifying, encapsulating, entrapping or lyophilizing. Pharmaceutical compositions may be formulated in a conventional manner using one or more physiologically acceptable carriers, which may be selected from excipients and auxiliaries that facilitate processing of the active compounds into preparations which can be used pharmaceutically.
The pharmaceutical compositions also may comprise suitable solid- or gel-phase carriers or excipients. Examples of such carriers or excipients include calcium carbonate, calcium phosphate, sugars, starches, cellulose derivatives, gelatin, and polymers such as polyethylene glycols. Thus, if a solid carrier is used, the preparation can be tableted, placed in a hard gelatin capsule in powder or pellet form or in the form of a troche or lozenge. The amount of solid carrier may vary, but generally will be from about 25 mg to about 1 g. If a liquid carrier is used, the preparation will be in the form of syrup, emulsion, soft gelatin capsule, sterile injectable solution or suspension in an ampule or vial or non-aqueous liquid suspension.
To obtain a stable water-soluble dose form, a pharmaceutically acceptable salt of an inventive agent is dissolved in an aqueous solution of an organic or inorganic acid, such as 0.3M solution of succinic acid or citric acid. If a soluble salt form is not available, the agent may be dissolved in a suitable cosolvent or combinations of cosolvents. Examples of suitable cosolvents include, but are not limited to, alcohol, propylene glycol, polyethylene glycol 300, polysorbate 80, gylcerin and the like in concentrations ranging from 0-60% of the total volume. In an exemplary embodiment, a compound of Formula I is dissolved in DMSO and diluted with water. The composition may also be in the form of a solution of a salt form of the active ingredient in an appropriate aqueous vehicle such as water or isotonic saline or dextrose solution.
A pharmaceutical carrier for hydrophobic compounds is a cosolvent system comprising benzyl alcohol, a nonpolar surfactant, a water-miscible organic polymer, and an aqueous phase. The cosolvent system may be a VPD co-solvent system. VPD is a solution of 3% w/v benzyl alcohol, 8% w/v of the nonpolar surfactant polysorbate 80, and 65% w/v polyethylene glycol 300, made up to volume in absolute ethanol. The VPD co-solvent system (VPD:5W) contains VPD diluted 1:1 with a 5% dextrose in water solution. This co-solvent system dissolves hydrophobic compounds well, and itself produces low toxicity upon systemic administration. Naturally, the proportions of a co-solvent system may be varied considerably without destroying its solubility and toxicity characteristics. Furthermore, the identity of the co-solvent components may be varied: for example, other low-toxicity nonpolar surfactants may be used instead of polysorbate 80; the fraction size of polyethylene glycol may be varied; other biocompatible polymers may replace polyethylene glycol, e.g. polyvinyl pyrrolidone; and other sugars or polysaccharides may be substituted for dextrose.
Alternatively, other delivery systems for hydrophobic pharmaceutical compounds may be employed. Liposomes and emulsions are known examples of delivery vehicles or carriers for hydrophobic drugs. Certain organic solvents such as dimethylsulfoxide also may be employed, although usually at the cost of greater toxicity. Additionally, the compounds may be delivered using a sustained-release system, such as semipermeable matrices of solid hydrophobic polymers containing the therapeutic agent. Various sustained-release materials have been established and are known by those skilled in the art. Sustained-release capsules may, depending on their chemical nature, release the compounds for a few weeks up to over 100 days. Depending on the chemical nature and the biological stability of the therapeutic reagent, additional strategies for protein stabilization may be employed.
It will be appreciated that the actual dosages of the agents used in the compositions of this invention will vary according to the particular complex being used, the particular composition formulated, the mode of administration and the particular site, host and disease being treated. Optimal dosages for a given set of conditions can be ascertained by those skilled in the art using conventional dosage-determination tests in view of the experimental data for an agent. For oral administration, an exemplary daily dose generally employed is from about 0.001 to about 1000 mg/kg of body weight, more preferably from about 0.001 to about 50 mg/kg body weight, and most preferably 1-20 mg/kg, with courses of treatment repeated at appropriate intervals. Administration of prodrugs are typically dosed at weight levels which are chemically equivalent to the weight levels of the fully active form. Intravenously, the most preferred doses will range from about 0.01 to about 10 mg/kg/minute during a constant rate infusion. Advantageously, the thrombin inhibitors may be administered in divided doses of two, three, or four times daily. Furthermore, they can be administered in intranasal form vial topical use of suitable intranasal vehicles, or via transdermal routes, using those forms of transdermal skin patches well known to those of ordinary skill in that art. To be administered in the form of a transdermal delivery system, the dosage administration will, or course, be continuous rather than intermittent throughout the dosage regime.
Therapeutically effective amounts of the agents of the invention may be used to treat diseases mediated by modulation or regulation of trypsin-like serine proteases. An xe2x80x9ceffective amountxe2x80x9d is intended to mean that amount of an agent that, when administered to a mammal in need of such treatment, is sufficient to effect treatment for a disease mediated by the activity of one or more trypsin-like serine proteases, such as thrombin. Thus, e.g., a therapeutically effective amount of a compound of the Formula I, salt, active metabolite or prodrug thereof is a quantity sufficient to modulate, regulate, or inhibit the activity of one or more protein kinases such that a disease condition which is mediated by that activity is reduced or alleviated.
The amount of a given agent that will correspond to such an amount will vary depending upon factors such as the particular compound, disease condition and its severity, the identity (e.g., weight) of the mammal in need of treatment, but can nevertheless be routinely determined by one skilled in the art. xe2x80x9cTreatingxe2x80x9d is intended to mean at least the mitigation of a disease condition in a mammal, such as a human, that is affected, at least in part, by the activity of one or more trypsin-like serine proteases, such as thrombin, and includes: preventing the disease condition from occurring in a mammal, particularly when the mammal is found to be predisposed to having the disease condition but has not yet been diagnosed as having it; modulating and/or inhibiting the disease condition; and/or alleviating the disease condition.
Proper formulation is dependent upon the route of administration chosen. The inventive compounds may be administered in intravenous (bolus or infusion), intraperitoneal, subcutaneous, or intramuscular form, all using forms well known to those of ordinary skill in the pharmaceutical arts. An effective but non-toxic amount of the compound desired can be employed as an anti-aggregation agent. For treating ocular build up of fibrin, the compounds may be administered intraocularly or topically as well as orally or parenterally.
For injection, the agents of the invention may be formulated into aqueous solutions, preferably in physiologically compatible buffers such as Hanks""s solution, Ringer""s solution, or physiological saline buffer. For transmucosal administration, penetrants appropriate to the barrier to be permeated are used in the formulation. Such penetrants are generally known in the art.
For oral administration, the compounds can be formulated readily by combining the active compounds with pharmaceutically acceptable carriers known in the art. Such carriers enable the compounds of the invention to be formulated as tablets, pills, dragees, capsules, liquids, gels, syrups, slurries, suspensions and the like, for oral ingestion by a patient to be treated. Pharmaceutical preparations for oral use can be obtained using a solid excipient in admixture with the active ingredient (agent), optionally grinding the resulting mixture, and processing the mixture of granules after adding suitable auxiliaries, if desired, to obtain tablets or dragee cores. Suitable excipients include fillers such as sugars, including lactose, sucrose, mannitol, or sorbitol; and cellulose preparations, for example, maize starch, wheat starch, rice starch, potato starch, gelatin, gum, methyl cellulose, hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, or polyvinylpyrrolidone (PVP). If desired, disintegrating agents may be added, such as crosslinked polyvinyl pyrrolidone, agar, or alginic acid or a salt thereof such as sodium alginate.
Dragee cores are provided with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, polyvinyl pyrrolidone, Carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquer solutions, and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablets or dragee coatings for identification or to characterize different combinations of active agents.
Pharmaceutical preparations which can be used orally include push-fit capsules made of gelatin, as well as soft, sealed capsules made of gelatin and a plasticizer, such as glycerol or sorbitol. The push-fit capsules can contain the active ingredients in admixture with fillers such as lactose, binders such as starches, and/or lubricants such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active agents may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin, or liquid polyethylene glycols. In addition, stabilizers may be added. All formulations for oral administration should be in dosages suitable for such administration. For buccal administration, the compositions may take the form of table ts or lozenges formulated in conventional manner.
For administration intranasally or by inhalation, the compounds for use according to the present invention are conveniently delivered in the form of an aerosol spray presentation from pressurized packs or a nebuliser, with the use of a suitable propellant, e.g., dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In the case of a pressurized aerosol the dosage unit may be determined by providing a valve to deliver a metered amount. Capsules and cartridges of gelatin for us e in an inhaler or insufflator and the like may be formulated containing a powder mix of the compound and a suitable powder base such as lactose or starch.
The compounds may be formulated for parenteral administration by injection, e.g., by bolus injection or continuous infusion. Formulations for injection may be present e d in unit-dosage form, e.g., in ampoules or in multi-dose containers, with an added preservative. The compositions may take such forms as suspensions, solutions or emulsions in oily or aqueous vehicles, and may contain formulatory agents such as su spending, stabilizing and/or dispersing agents.
Pharmaceutical formulations for parenteral administration include aqueous solutions of the active compounds in water-soluble form. Additionally, suspensions of the active agents may be prepared as appropriate oily injection suspensions. Suitable lipophilic solvents or vehicles include fatty oils such as sesame oil, or synthetic fatty acid esters, such as ethyl oleate or triglycerides, or liposomes. Aqueous injection suspensions may contain substances which increase the viscosity of the suspension, such as sodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, the suspension may also contain suitable stabilizers or agents which increase the solubility of the compounds to allow for the preparation of highly concentrated solutions.
Alternatively, the active ingredient may be in powder form for constitution with a suitable vehicle, e.g., sterile pyrogen-free water, before use. The compounds may also be formulated in rectal compositions such as suppositories or retention enemas, e.g., containing conventional suppository bases such as cocoa butter or other glycerides.
In addition to the formulations described above, the compounds may also be formulated as a depot preparation. Such long-acting formulations may be administered by implantation (for example, subcutaneously or intramuscularly) or by intramuscular injection. Thus, for example, the compounds may be formulated with suitable polymeric or hydrophobic materials (for example, as an emulsion in an acceptable oil) or ion-exchange resins, or as sparingly soluble derivatives, for example, as a sparingly soluble salt.
The thrombin inhibitors can also be co-administered with suitable anti-coagulation agents or thrombolytic agents such as plasminogen activators or streptokinase to achieve synergistic effects in the treatment of various ascular pathologies. For example, thrombin inhibitors enhance the efficiency of tissue plasminogen activator-mediated thrombolytic reperfusion. Thrombin inhibitors may be administered first following thrombus formation, and tissue plasminogen activator or other plasminogen activator is administered thereafter. They may also be combined with heparin, aspirin, or warfarin.
Anticoagulant therapy is indicated for the treatment and prevention of a variety of thrombotic conditions, particularly coronary artery and cerebrovascular disease. Those experienced in this field are readily aware of the circumstances requiring anticoagulant therapy. The term xe2x80x9cpatientxe2x80x9d used herein is taken to mean mammals such as primates, including humans, sheep, horses, cattle, pigs, dogs, cats, rats, and mice.
The dosage regimen utilizing the thrombin inhibitors is selected in accordance with a variety of factors including type, species, age, weight, sex and medical condition of the patient; the severity of the condition to be treated; the route of administration; the renal and hepatic function of the patient; and the particular compound or salt thereof employed. An ordinarily skilled physician or veterinarian can readily determine and prescribe the effective amount of the drug required to prevent, counter, or arrest the progress of the condition.
Thrombin inhibition is useful not only in the anticoagulant therapy of individuals having thrombotic conditions, but is useful whenever inhibition of blood coagulation is required such as to prevent coagulation of stored whole blood and to prevent coagulation in other biological samples for testing or storage. Thus, the thrombin inhibitors can be added to or contacted with any medium containing or suspected of containing thrombin and in which it is desired that blood coagulation be inhibited, e.g. when contacting the mammal""s blood with material selected from the group consisting of vascular grafts, stents, orthopedic prothesis, cardiac prosthesis, and extracorporeal circulation systems.
The inventive agents may be prepared using the reaction routes and synthesis schemes as described below, employing the techniques available in the art using starting materials that are readily available.
The preparation of preferred compounds of the present invention is described in detail in the following examples, but the artisan will recognize that the chemical reactions described may be readily adapted to prepare a number of other protein kinase inhibitors of the invention. For example, the synthesis of non-exemplified compounds according to the invention may be successfully perfomed by modifications apparent to those skilled in the art, e.g. by appropriately protecting interfering groups, by changing to other suitable reagents known in the art, or by making routine modifications of reaction conditions. Alternatively, other reactions disclosed herein or known in the art will be recognized as having applicability for preparing other compounds of the invention.